Survival and success rates of soft-milled cobalt-chromium-ceramic full-arch screw-retained implant-supported prostheses: a 2- to 7-year follow-up retrospective study.

OBJECTIVES: To retrospectively assess implant and prosthesis survival and success and patient satisfaction in edentulous patients treated with soft-milled cobalt-chromium-ceramic full-arch screw-retained implant-supported prostheses (SCCSIPs) over a mean clinical follow-up period of 4 years. MATERIALS AND METHODS: Twenty-one edentulous patients, aged 47-80 years, underwent restoration with 27 SCCSIPs (210 implants, 349 crown units). After definitive prosthesis insertion, patients participated in a yearly dental check-up recall program, including clinical and radiographic examinations. All supporting implants were examined for biological and technical complications, and the restorations were evaluated using modified California Dental Association (CDA) criteria. Life table and Kaplan-Meier survival analyses were performed. Patient satisfaction regarding chewing, esthetics, comfort, and phonetics was evaluated using a visual analog scale (VAS). RESULTS: After a mean of 4 years, two implants (3.8%) failed, with no effect on prosthesis survival. The cumulative survival rate was 98.6% and 100% for implants and prostheses, respectively. The most frequent minor biological complication was soft tissue recession (11%). Porcelain fracture was the only major technical complication (0.6% of crown units), while the only minor technical complication was porcelain chipping (4% of crown units), which required only polishing. Overall, 66.7% of the prostheses were free of technical complications. Patient satisfaction was high to very high. Less satisfaction was noted with comfort due to soft tissue recession and food impaction. CONCLUSIONS: Within the limitations of this study, we conclude that the survival and success rates of SCCSIPs in edentulous patients were excellent. CLINICAL RELEVANCE: SCCSIPs should be considered when planning metal-ceramic implant-supported restorations.

Safety, tolerability and efficacy of a novel self-use biodegradable device for management of obesity.

OBJECTIVE: Obesity is a major public health issue with significant impact on quality of life, morbidity and mortality rates. It is estimated that if the current trends continue, 18% of men and 21% of women worldwide will be obese by 2025. All the current therapies are not optimal due to limited efficacy or safety; thus, there is a need for additional devices for the treatment of obesity. This study aimed to examine the safety, tolerability, and efficacy of a biodegradable encapsulated Epitomee device for weight loss. The technology is based on absorbent pharmaceuticals polymers and bonding materials that self-expand in the stomach to create a pH-sensitive super absorbent gel structure for weight loss. METHODS: A prospective, 12-week twice daily use of the encapsulated device in patients with body mass index of 27-40 kg m-2. Efficacy endpoints were the percent total body weight loss (%TBWL), proportion of participants with 5% TBWL and changes in cardio-metabolic markers. Safety analysis included evaluation of adverse events, laboratory and endoscopic findings. RESULTS: Overall, 52 patients completed the study. TBWL per intension-to-treat analysis was 3.68 ± 3.07% (3.23 ± 2.69 kg) and 4.52 ± 2.97% (3.95 ± 2.57 kg) per protocol. No device serious adverse effects reported. The most common adverse events were headache (18.1%), viral infection (11.5%), abdominal discomfort (10.1%), bloating (7.9%), nausea and constipation (5% each) and flatulence (4.3%). Endoscopy in 26 patients revealed mild, asymptomatic gastric/duodenal erythema without erosions in five patients. CONCLUSIONS: Twelve weeks of Epitomee capsules treatment combined with lifestyle counselling resulted in 3.68-4.52% of TBWL. With continued research, the Epitomee capsules have considerable potential to become a non-invasive, safe and effective treatment option for weight loss.

Complete rehabilitation of patients with bruxism by veneered and non-veneered zirconia restorations with an increased vertical dimension of occlusion: an observational case-series study.

PURPOSE: To assess the clinical performance of the complete rehabilitation of a series of patients with bruxism treated by teeth- and implant-supported veneered and non-veneered zirconia restorations with an increased vertical dimension of occlusion (VDO). METHODS: Ten patients with bruxism, aged 62-70 years, were treated with 108 veneered and 142 non-veneered zirconia restorations and attended the recall appointment. The mean observation period was 28.2 (±16.8) months. The patients were identified from records, and clinical details were retrieved from their files. In the recall appointment, the restorations were evaluated using modified California Dental Association (CDA) criteria. The periodontal probing depth, bleeding index, presence of caries and implant survival and success rate were recorded. RESULTS: No biological complications were recorded for any restorations. The success and survival rate of all implants was 100%. The overall mean survival and success rate of all restorations was 99.6%. In the veneered group, the predominant complication was minor veneer chipping (13.9%) on the incisal edge, which required only polishing (grade 1); in the non-veneered group, the predominant failure was open proximal contacts between the implant restoration and adjacent teeth (9%); only one implant restoration needed repair. One restoration was replaced due to a horizontal tooth fracture. CONCLUSIONS: Within the limitations of this study, we conclude that the survival and success rate of monolithic zirconia restorations installed in patients with bruxism was excellent, although the veneered zirconia restorations showed a high rate of minor veneer chipping, which required only polishing.

The effect of gingival wall location on the marginal seal of class ii restorations prepared with a flowable bulk-fill resin-based composite.

SureFil SDR is a flowable resin-based composite that allows a single incremental bulk placement. The marginal seal of SureFil SDR at the gingival margins of class II restorations located apical to the cemento-enamel-junction (CEJ) has not been adequately evaluated compared to those located occlusal to the CEJ. Forty class II cavities were prepared in human molars. The gingival margins of 20 preparations were located 0.5 mm occlusal to the CEJ, and the other 20 preparations were located 0.5 mm apical to the CEJ. The cavities surfaces were bonded with XenoV dental adhesive and filled with SDR in one bulk increment up to 4 mm, after which they were covered with CeramX. The teeth were subjected to thermo-and load-cycling, and their gingival margins were exposed to 0.5% basic-fuchsin solution. The specimens were sectioned mesio-distally and scored for microleakage. A Wilcoxon test for pairwise comparison was performed to determine significance. Dye penetration was observed in 30% of the 20 restorations with cavo-surface margins located occlusal to the CEJ and in 55% of the 20 restorations with cavo-surface margins located apical to the CEJ. The bulk-fill flowable resin base SureFil SDR with XenoV dental adhesive provided a better marginal seal in class II restorations with gingival margins above the CEJ compared to restorations with gingival margins below the CEJ. SDR should not be recommended for class II cavity preparations with gingival margins located below the CEJ.

Survey results of currently used techniques and materials by dentists for fabrication of complete removable dental prostheses.

The present study was based on the screening of procedures and materials used by dentists in constructing complete removable dental prostheses, considering their professional background and years of experience. Questions arose regarding differences and similarities within the local dental society, of which approximately 75% were trained in Europeans countries with differences in academic background, and compared to international dentistry. A questionnaire web site was used in order to anonymously collect answers to the survey by general practicing and specialist dentists. The survey contained 28 multiple choice questions that were based on 5 parameters: years of experience, location of dental studies, specialization, location and field of specialization. One hundred one survey sheets were collected. The results show statistical significance (p less than 0.05) for the following comparisons: practicing time had an impact on practitioner-technician interaction, while the level of qualification had an effect on articulator and face-bow usage. Location of dental studies, specialization and residency had impact on final impression materials, teeth set-up as well as tooth shape and color selection. The differences found between dentists’ working techniques set the ground for future research that should focus on rating the success of the different methods towards a goal of creating recommendations for an ideal set of working techniques.

The effect of enriching denture adhesives with chlorhexidine diacetate on the proliferation of Candida albicans: an in vitro analysis.

Denture adhesives are widely used in dentistry. The growth of Candida albicans on the denture base is common on both jaws, particularly in elderly patients. A strong correlation has been found between the growth of Candida albicans and dentures, potentially leading to denture stomatitis. Recent studies show that the addition of chlorhexidine diacetate (CHDA) to denture adhesives can have a protective effect against denture stomatitis. The aim of this research is to examine the antifungal properties of three denture adhesives when CHDA is added. We hypothesize that the addition of CHDA to denture adhesives will result in antifungal activity against Candida albicans. Three denture adhesive materials (Corega, Protefix and Kukident) were prepared with and without the addition of CHDA in order to measure whether CHDA increased the resistance of the adhesives to the growth of Candida albicans. Dynamic and static intercellular changes were assessed through direct contact test (DCT) and confocal microscopy. The tested materials were evaluated immediately after setting and at 14 days post-setting. When combined with CHDA, all three denture adhesive materials decreased Candida albicans growth compared to the control (p less than 0.0001). This inhibition of growth was more prominent in the first 24 h. The confocal microscopy results supported the DCT findings. The addition of CHDA to denture adhesive pastes inhibits growth of Candida albicans, and may potentially reduce the incidence of denture stomatitis. Further studies are warranted.

Temperature changes of one-piece implants during the setting of acrylic resin temporary crown. The effect of implant diameter. An in vitro study.

The purpose of this work is to evaluate changes in temperature of one-piece titanium implant surface during the setting of acrylic resin temporary crowns and to correlate thermal changes to implant diameter. Thirty-three one-piece implants (ARRP, Alpha-Biotec) were divided into 3 groups according to diameter size (G1=3 mm, G2=3.3 mm, G3=3.6 mm). Implants were mounted on an acrylic glass apparatus. Thermocouples were positioned at the most coronal thread. Lower incisor temporary polycarbonate crowns were filled with 80 µL of self-curing acrylic resin and positioned immediately on the implant abutment. Thermal changes of the implant surface were recorded continuously for 10 min. Data were statistically analyzed using one-way analysis of variance. The mean initial temperature (C0) of groups G1, G2 and G3 was similar (24.79±0.78ºC, 25.26±0.63ºC, 24.97±1.06ºC, respectively). The setting of the acrylic resin temporary crown resulted in a significant increase in the implant surface temperature of all groups. The mean thermal amplitude (ΔC) for groups G1, G2 and G3 were 6.79±1.02ºC, 6.61±0.94ºC, 6.65±1.26ºC, respectively. The mean time to maximum temperature (Tmax) for groups G1, G2 and G3 were 337.38±42.91 sec, 324.69±41.46 sec and 317.98±37.91 sec respectively (P>0.05). Direct application of auto-polymerizing resin to the titanium abutment of one-piece implants significantly increased the cervical implant surface temperature. Implant diameter did not influence the temperature changes.

Prognostic significance of supradiaphragmatic lymph nodes at initial presentation in patients with stage III high-grade serous ovarian cancer.

PURPOSE: To identify the optimal size threshold and to assess the prognostic significance of supradiaphragmatic lymph nodes at initial presentation of patients with high-grade serous ovarian cancer (HGSC). METHODS: This IRB-approved, HIPAA-compliant retrospective study included baseline pretreatment staging abdominal CTs of 88 women (mean age 62 years, SD 10.4, range 29-85) with FIGO stage III HGSC. Patients with stage IV disease were excluded due to worse prognosis and management guided by distant metastases. Two fellowship-trained radiologists independently reviewed abdominal CTs to record the presence of supradiaphragmatic nodes, abdominal lymphadenopathy, peritoneal carcinomatosis, and ovarian mass. Progression-free survival (PFS) and overall survival (OS) were recorded after median 79 months follow-up (IQR 58-115, range 13-144). The optimal short-axis size threshold for supradiaphragmatic lymphadenopathy was determined by correlating 3, 4, 5, 6, 7, and 10 mm thresholds with PFS and OS using Log-rank test. Prognostic significance of supradiaphragmatic lymphadenopathy was assessed using Cox proportional hazards models. RESULTS: There was good interobserver agreement for presence (κ = 0.65, 95%CI 0.51-0.79) and size (ICC = 0.77, 95%CI 0.66-0.86) of supradiaphragmatic nodes. 5 mm short-axis size threshold was associated with significantly shorter PFS (median 14 months, IQR 11-17 vs. 23 months, IQR 12-59; p = 0.02) and OS (median 44 months, IQR 27-69 vs. 65 months, IQR 45-96; p = 0.03). Total 38/88 (43%) patients had supradiaphragmatic lymphadenopathy. On Cox proportion hazards analysis, supradiaphragmatic lymphadenopathy was significantly associated with shorter PFS (p = 0.02; HR 1.81, 95%CI 1.11-2.96) and OS (p = 0.008; HR 2.11, 95%CI 1.21-3.65). CONCLUSION: In patients with stage III HGSC, supradiaphragmatic lymphadenopathy is associated with shorter PFS and OS. Further studies would help determine its implications on staging, decision regarding neoadjuvant therapy, and surgical technique.

Efficacy of XP-endo finisher files in the removal of calcium hydroxide paste from artificial standardized grooves in the apical third of oval root canals.

AIM: To compare the efficacy of the XP-endo finisher file (XP) (FKG Dentaire, La Chaux de Fonds, Switzerland) to that of passive ultrasonic irrigation (PUI) and conventional syringe and needle irrigation (SNI) in the removal of calcium hydroxide paste from an artificial standardized groove in the apical third of root canals. METHODOLOGY: The root canals of 68 mandibular incisors with single oval canals were prepared using Mtwo instruments (VDW GmbH, Munich, Germany) up to size 40, .04 taper. Each tooth was split longitudinally, and in one half of the root, a standardized groove was prepared in the apical part of the specimen. The grooves were filled with Ca(OH)2 , and the root halves were reassembled. The roots were randomly divided into two control groups (n = 4) and three experimental groups (n = 20) according to the Ca(OH)2 methods used: XP, PUI and SNI. The amount of remaining medicament was evaluated under X25 magnification using a 4-grade scoring system. Kappa values were calculated for intra- and interobserver agreement evaluation. The differences in the Ca(OH)2 scores amongst the different groups were analysed using the Kruskal-Wallis test. RESULTS: None of the tested methods could completely clean the Ca(OH)2 from the artificial standardized groove in the apical third of the root canals. XP and PUI removed significantly more Ca(OH)2 than SNI (P < 0.001), with no significant differences between them (P = 0.238). CONCLUSIONS: XP and PUI were more effective in removing Ca(OH)2 from artificial standardized grooves in the apical third of root canals than SNI.

Mesenchymal stem cells secretomes' affect multiple myeloma translation initiation.

Bone marrow mesenchymal stem cells' (BM-MSCs) role in multiple myeloma (MM) pathogenesis is recognized. Recently, we have published that co-culture of MM cell lines with BM-MSCs results in mutual modulation of phenotype and proteome (via translation initiation (TI) factors eIF4E/eIF4GI) and that there are differences between normal donor BM-MSCs (ND-MSCs) and MM BM-MSCs (MM-MSCs) in this crosstalk. Here, we aimed to assess the involvement of soluble BM-MSCs' (ND, MM) components, more easily targeted, in manipulation of MM cell lines phenotype and TI with specific focus on microvesicles (MVs) capable of transferring critical biological material. We applied ND and MM-MSCs 72h secretomes to MM cell lines (U266 and ARP-1) for 12-72h and then assayed the cells' (viability, cell count, cell death, proliferation, cell cycle, autophagy) and TI (factors: eIF4E, teIF4GI; regulators: mTOR, MNK1/2, 4EBP; targets: cyclin D1, NFκB, SMAD5, cMyc, HIF1α). Furthermore, we dissected the secretome into >100kDa and <100kDa fractions and repeated the experiments. Finally, MVs were isolated from the ND and MM-MSCs secretomes and applied to MM cell lines. Phenotype and TI were assessed. Secretomes of BM-MSCs (ND, MM) significantly stimulated MM cell lines' TI, autophagy and proliferation. The dissected secretome yielded different effects on MM cell lines phenotype and TI according to fraction (>100kDa- repressed; <100kDa- stimulated) but with no association to source (ND, MM). Finally, in analyses of MVs extracted from BM-MSCs (ND, MM) we witnessed differences in accordance with source: ND-MSCs MVs inhibited proliferation, autophagy and TI whereas MM-MSCs MVs stimulated them. These observations highlight the very complex communication between MM and BM-MSCs and underscore its significance to major processes in the malignant cells. Studies into the influential MVs cargo are underway and expected to uncover targetable signals in the regulation of the TI/proliferation/autophagy cascade.

First trimester human placental factors induce breast cancer cell autophagy.

Placental factors, progesterone included, facilitate breast cancer cell line (BCCL) motility and thus may contribute to the advanced breast cancer found during pregnancy. Cancer and placental implantations are similar; the last is accompanied by extravillous trophoblast cell invasion and autophagy which are interlinked. We aimed to analyze the effect of first trimester human placenta on BCCL autophagy. BCCLs (MCF-7/T47D) were cultured with placental explants (60 h) or placental supernatants (24 h). Following cultures, BCCLs were sorted out for RNA/protein extraction. RNA served for microarray/qPCR (BNIP3) and protein for Western blot (HIF1α, LC3BII) analyses. Inhibitors were added to the placenta-MCF-7 coculture or placental supernatants (autophagy inhibitor-3MA, progesterone receptor (PR) inhibitor-RU486, and HIF1α inhibitor-Vitexin) in order to evaluate their effects on BCCL motility and LC3BII/HIF1α expression. LC3BII (an autophagy marker) expression was elevated in BCCLs following placental explant coculture and exposure to placental supernatants. The autophagy inhibitor (3MA) repressed the placenta-induced MCF-7/T47D migration, establishing a connection between BCCL autophagy and migration. Microarray analysis of MCF-7 following placenta-MCF-7 coculture showed that "HIF1α pathway," a known autophagy facilitator, was significantly manipulated. Indeed, placental factors elevated HIF1α and its target BNIP3 in the BCCLs, verifying array results. Lastly, PR inhibitor reduced HIF1α expression and both PR and HIF1α inhibitors reduced MCF-7 LC3BII expression and motility, suggesting involvement of the PR-HIF1α axis in the autophagy process. Placental factors induced BCCL autophagy that is interlinked to their motility. This suggests that autophagy-related molecules may serve as targets for therapy in pregnancy-associated breast cancer.

The association between dental wear and reduced vertical dimension of the face: a morphologic study on human skulls.

OBJECTIVE: The aim of this study was to explore the relationship between dental wear and facial morphology, with particular reference to the occlusal vertical dimension, in modern human skulls. DESIGN: One hundred and three skulls (52 men and 51 women) between the ages of 20 and 50+ years old were studied. The selected skulls were from a modern period (the 17th and the 18th centuries) and included at least one entire condyle and had at least 3 posterior teeth (premolar or molar) in each quadrant to allow for dental articulation. Occlusal wear was evaluated using ordinal scale (0-4) and vertical occlusal dimension was evaluated by measuring upper facial height (UFH), lower facial height (LFH), LFH-to-UFH ratio (L-U-R) and dental wear. Based on the occlusal wear score, two groups were defined: with and without significant wear. RESULTS: Significant relation was observed between age and dental wear (P<0.01). No significant differences were found in the LFH (P=0.847) or UFH (P=0.108) between the two wear groups. In addition, no significant difference (P=0.132) was demonstrated in the LFH-to-UFH ratio between the groups. No difference was observed in the dental wear score between genders (P=0.321). CONCLUSION: Within its limitations, this study demonstrated that dental wear does not influence the vertical dimension of occlusion. Our assumption is that the dento-facial complex fully compensates for the dental effects of wear throughout life.

The effects of in vitro fluoride mouth rinse on the antibacterial properties of orthodontic cements.

OBJECTIVES: To investigate the ability of orthodontic cements to regain their antibacterial effect after aging for 1 month, followed by 2 weeks of fluoride 'recharging' through daily fluoride rinse. MATERIALS AND METHODS: Four orthodontic cements were tested: composite resin-based materials (Transbond XT and Transbond Plus), a conventional glass ionomer cement (CX Plus) and dual-cured resin-reinforced glass ionomer cement (Fuji ORTHO LC) by direct contact test. After polymerization and a 30-day aging process, the samples were rinsed daily with 0.05% NaF solution for 14 days. Twenty-four hours after the last fluoride rinse, Streptococcus mutans cells (approximately 1 x 10(6) ) were placed on the surface of each sample for 1 h at 37°C to establish direct contact. Bacterial growth was monitored for 24 h by temperature-controlled spectrophotometry. Similar experiments were conducted after aging for 48 h and 72 h after the last fluoride rinse. One-way anova, two-way anova, and Tukey's multiple comparison test were applied to the data. RESULTS: Twenty-four hours after the last fluoride rinse, the resin-modified glass ionomer and the glass ionomer showed potent antibacterial properties, whereas the composite-based material Transbond XT encouraged bacterial growth. After 72 h, only Fuji Ortho LC maintained its bacteriostatic properties while all the other tested materials showed no antibacterial activity. CONCLUSIONS: Fourteen days daily fluoride rinse temporarily revives the antibacterial properties of the resin-modified glass ionomer cement and glass-ionomer-based cement.

The effect of heat shock protein 27 on extravillous trophoblast differentiation and on eukaryotic translation initiation factor 4E expression.

Heat shock protein (HSP27) is expressed in human placentae. Previously, we showed that HSP27 is expressed in the villous cell column of first trimester placental explants and in extravillous trophoblast (EVT) cells. EVT differentiation is accompanied by increased motility, matrix metalloproteinase (MMP) activity, decreased proliferation and expression of specific markers such as HLAG and CD9. HSP27 regulates cell apoptosis, migration, protein stability and the availability of eukaryotic translation initiation factors, such as eukaryotic translation initiation factor 4E (eIF4E). eIF4E supports trophoblast cell proliferation and survival. We wanted to explore the effect of HSP27 silencing on trophoblast cell phenotype, EVT markers and eIF4E expression and regulators [4E-binding protein (4E-BP1) and MAP kinase-interacting kinase (MNK1)]. This study evaluated the effect of HSP27 siRNA on placental explant and HTR-8/SVneo migration, MMP activity/mRNA, cell death, cell cycle, HLAG/CD9 levels, and eIF4E and its regulators' total and phosphorylated levels. Furthermore, we evaluated HSP27 levels in placentae exposed to ribavirin, which triggers EVT differentiation. We found that HSP27 silencing increased cell death in HTR-8/SVneo and placental explants. Furthermore, it reduced HTR-8/SVneo migration and EVT outgrowth from the explants (P < 0.05), MMP2 activity and expression of EVT markers HLAG and CD9 (in placental explants and HTR-8/SVneo, respectively, P < 0.05). Induction of EVT differentiation by ribavirin elevated HSP27 levels. Finally, HSP27 silencing in both HTR-8/SVneo and placental explants reduced eIF4E levels (33 and 28%, respectively, P < 0.05) and the levels of its regulators 4E-BP1 and MNK1 (37 and 32%, respectively, done on HTR-8/SVneo only), but not their phosphorylated forms. Altogether, our results suggest that HSP27 contributes to EVT cell differentiation.

In vitro CPC retention and VSC adsorption by IPM oil droplets: possible mechanisms of action of a two phase mouthwash.

Two phase oil-water mouthwash has been previously shown to efficiently bind oral microorganisms, relying on their cell surface hydrophobicity. The aim of the present in vitro study was to test the cetylpyridinium chloride (CPC) retention and volatile sulfide compounds (VSCs) adsorption abilities of the oil droplets created by mixing of a two phase oil-water solution. VSC adsorption was assayed using a salivary incubation assay and garlic powder solutions, and demonstrated using microscopic sulfide assay. CPC retention was assayed by kinetic and endpoint measurement of Streptococcus salivarius outgrowth using microplate (ELISA) reader. Results showed that the isopropyl myristate (IPM) oil droplets in the two phase solutions were able to adsorb 68-80% of VSCs. CPC at a concentration of 0.05% was most affectively retained by the oil droplets showing a significantly increase in residual antibacterial activity against Streptococcus salivarius. These results taken together, suggests that VSC adsorption and CPC retention by IPM oil droplets may be two additional mechanisms in the activity of the two phase mouthwash formulation.

The involvement of eukaryotic translation initiation factor 4E in extravillous trophoblast cell function.

UNLABELLED: Extravillous trophoblast cells (EVT) are major players in placental implantation. They differentiate in the villous cell column, invade to the uterus and remodel the uterine spiral arteries. Trophoblast and tumor cells have similar invasion mechanisms, share similar biochemical mediators (e.g. c-myc, MMP9) and growth-factors (e.g. VEGF). The mRNA of these proteins has extremely structured 5-UTR and their translation is highly dependent on eukaryotic-translation-initiation-factor-4E (eIF4E). Cancer cells have elevated eIF4E and are more vulnerable to its silencing than normal cells. We speculated that like cancer, trophoblast function is highly eIF4E dependent. OBJECTIVE: Analyze eIF4E involvement in EVT differentiation and function. STUDY DESIGN: EIF4E levels were assessed in first-trimester human placentae and in placental explants before and after EVT differentiation. The effect of eIF4E knockdown (siRNA, ribavirin) on the phenotype of placental explant and EVT cell lines (HTR-8/SVNEO) was evaluated. Tested parameters included eIF4E and its target levels, migration, invasion, cell death, cell cycle and cell count. RESULTS: High eIF4E levels were found in cytotrophoblast and especially EVT cells during their differentiation in the villi, compared to other placental cell types. EIF4E silencing increased cell death and cell cycle arrest in placental explants and HTR-8/SVNEO cells. Although it induced EVT outgrowth in the placental explants, it reduced HTR-8/SVNEO motility, reflecting the importance of using ex vivo models that include an intact placental microenvironment in its original architecture. CONCLUSIONS: Our results suggest that eIF4E prevents final EVT differentiation and supports placental cell proliferation and survival. A balance between cell proliferation and differentiation is crucial for placental development and implantation.

The efficacy of five techniques for removing root filling material: microscopic versus radiographic evaluation.

AIM: To test and compare the efficacy of five methods for the removal of root filling material and to test the hypothesis that radiographs fail to represent the real extent of remaining material on canal walls. METHODOLOGY: Fifty maxillary anterior single-rooted teeth with straight root canals were selected. The coronal third of each root canal was prepared with Gates-Glidden drills to number 3, whilst the apical two-thirds were prepared with manual K-files to size 40. Root fillings were performed using lateral compaction with gutta-percha and AH-26. After full setting, the coronal third of the root filling was removed with Gates-Glidden drills and the teeth divided into five groups (n=10). The remaining root filling material was then removed with either Hedström files and chloroform (25 µL), using size 40 as the last file, SafeSider files, using a NiTi Pleezer reamer with a 0.06 taper followed by size 40 reciprocating file, with or without chloroform, or ProTaper Universal retreatment files (D2, D3) with or without chloroform. Reaching working length with no more gutta-percha on the last file was defined as the endpoint for all procedures. The presence of remaining filling material was first evaluated radiographically and then by the microscopic evaluation of split roots. The time required to accomplish the procedure was also recorded. anova and anova with repeated measures were used for statistical analysis of the results. RESULTS: Overall, 11-26% of the canal wall remained covered with filling material; no significant difference was found between the groups. The mechanized methods were faster than manual removal of filling material (P < 0.01); the use of solvent did not speed up the mechanized procedures. Radiographic evaluation failed to adequately and reliably detect the extent of filling material remaining on the canal walls, which was later observed by microscopic evaluation. CONCLUSIONS: All methods left root canal filling material on the canal walls. Radiographic evaluation failed to detect the extent of remaining root filling material, which could only be detected using microscopy.

Hormone-dependent placental manipulation of breast cancer cell migration.

BACKGROUND: Breast cancer during pregnancy is often more advanced than in non-pregnant women. Nevertheless, no case of metastasis inside the placenta has been reported. Previously, we showed that placental-explants eliminated breast cancer cells from their surroundings, due to cell-death and elevated migration. Our objective was to find the underlying mechanisms of these phenomena. METHODS AND RESULTS: Our model contained Michigan Cancer Foundation 7 (MCF7) or T47D cells co-cultured with and without human placental explants. Microarray analysis, validated by quantitative PCR, of MCF7 following their placental co-culture suggested activation of estrogen (E(2)) signaling. As extensive cross-talk exists between E(2) and progesterone, their involvement in mediating placental effects on breast cancer cells was tested. Indeed, addition of E(2) and progesterone receptor (ER and PR) inhibitors to the co-culture system reduced cancer cell motility, yet did not alter cell-cycle or death. E(2) and progesterone concentrations in placental media were found to be similar to those of early pregnancy blood levels. Interestingly, placental-breast cancer co-culture media contained lower progesterone (P < 0.05) and higher E(2) (200%, P < 0.05) levels than placentae cultured separately. Placental supernatant and E(2) and progesterone at placental levels were sufficient to increase MCF7 and T47D migration and invasion (P < 0.05), yet did not alter MCF7 cell-cycle or death. Furthermore, placental supernatant elevated p38 and Jun N-terminal kinase (JNK) phosphorylation in both cell lines (P < 0.05). Inhibitors of JNK, ER and PR reversed MCF7 and T47D motility induced by the placenta, suggesting their involvement. CONCLUSIONS: We suggest that E(2) and progesterone contribute to cell migration away from placental areas. We hypothesize that they may increase metastatic spread to other organs in pregnancy.

Breast cancer characteristics are modified by first trimester human placenta: in vitro co-culture study.

BACKGROUND: Pregnant women with breast cancer present with a more advanced disease compared with non-pregnant women. Nevertheless, breast cancer metastasis to the placenta is rare. Trophoblast/tumor implantations share the same biochemical mediators, while only the first is stringently controlled. We hypothesized that the same mechanisms that affect/restrain placental implantation may inhibit metastatic growth in the placenta. We aimed to analyze the effects of human placenta on breast cancer cells. METHODS: First trimester human placental explants were co-cultured with MCF-7/T47D-eGFP tagged cells. Following culture, placenta/cancer cells/both were fixed, paraffin embedded and sliced for immunohistochemical analysis or sorted by their eGFP expression for future analysis. The tested parameters were: proliferation (immunohistochemistry)/cell cycle (FACS), apoptosis (immunohistochemistry/FACS), cell count/adhesion/distribution around the placenta (cell sorter, visual observation and counting), matrix metalloproteinase activity (zymogram) and estrogen receptor (ER) expression (western blotting, immunohistochemistry). RESULTS: Reduced breast cancer cell numbers (45%↓, 48%↓ for MCF-7/T47D, respectively, P < 0.05) were observed near the placenta. The placenta elevated MCF-7 sub-G1 phase and modestly elevated apoptosis (3-17%↑ for T47D/MCF-7, respectively, P < 0.05). Our findings demonstrate breast cancer cell migration from the placenta as: (i) T47D/MCF-7 cells changed their morphology to that of motile cells; (ii) elevated MMPs activity was found in the co-culture; (iii) placental soluble factors detached breast cancer cells; and (4) the placenta reduced MCF-7/T47D cells' ER expression (a characteristic of motile cells). CONCLUSIONS: MCF-7/T47D cells are eliminated from the placental surroundings. Analyzing the causes of these phenomena may suggest biological pathways for this event and raise new therapeutic targets.